Abstract：
:Recombinant proteins are routinely expressed in heterologous expression systems such as human embryonic kidney 293 (HEK 293) cells. The efficiency of the expression is critical when the expressed protein must be characterized at the single-cell level. Here we describe a simple method by which the protein expression efficiency in single HEK 293 cells is enhanced by coexpressing simian virus 40 large T antigen (TAg), a powerful oncoprotein. Using the GluR2 ionotropic glutamate receptor as an example, we found that the receptor expression in single HEK 293S cells increased approximately seven-fold. The ratio of the plasmid amount of TAg to that of the receptor was optimized at 1:10, while the receptor function was unaffected in the presence of TAg. We further used fluorescence imaging from a population of cells as an independent detection method and found a similar increase in expression of green fluorescent protein (GFP) by TAg coexpression. This method is thus applicable for enhancing the expression of both membrane and soluble proteins at the single-cell level. More importantly, the function of a protein can be studied directly in intact cells, a feature particularly useful for studying membrane proteins.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Huang Z,Li G,Pei W,Sosa LA,Niu Ldoi
10.1016/j.jneumeth.2004.09.009subject
Has Abstractpub_date
2005-03-15 00:00:00pages
159-66issue
1eissn
0165-0270issn
1872-678Xpii
S0165-0270(04)00329-2journal_volume
142pub_type
杂志文章abstract：:We describe a method for in vivo confocal fluorescence imaging of synaptic terminals and subsequent electron microscopic reconstructions of the same terminals. By iontophoretically applying lipophilic dye to nerve terminals at a single neuromuscular junction with a sharp microelectrode in living neonatal mice, we were...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(99)00096-5
更新日期：1999-10-30 00:00:00
abstract：:MicroRNAs (miRNAs) finely tune messenger RNA (mRNA) expression. As the brain is a highly heterogeneous tissue, physiologically relevant miRNA expression profiling greatly benefits from sampling brain regions or nuclei. MiRNA expression profiling from individual samples is also important for investigating potential dif...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2012.05.033
更新日期：2012-07-30 00:00:00
abstract：:The concentration of free calcium within the lumen of the endoplasmic reticulum ([Ca2+]L) fluctuates between 100 and 1000 microM. High [Ca2+]L provides an electro-driving force for Ca2+ release and supports high Ca2+ diffusion rate within the endoplasmic reticulum lumen. Fluctuations in [Ca2+]L also regulate numerous ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章,评审
doi：10.1016/s0165-0270(02)00300-x
更新日期：2002-12-31 00:00:00
abstract：:Prepulse inhibition (PPI) of the acoustic startle response is a measure of sensory motor gating, and is affected in various neuropsychiatric disorders. Although PPI has been used extensively to study both the neural effects of such conditions, as well as in the search for animal models, a number of critical issues hav...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2008.06.014
更新日期：2008-08-30 00:00:00
abstract：:We investigate the mode locking properties of simple dynamical models of pulse-coupled neurons to two tones, i.e., simple musical intervals. A recently proposed nonlinear synchronization theory of musical consonance links the subjective ranking from consonant to dissonant intervals to the universal ordering of robustn...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2009.06.041
更新日期：2009-09-30 00:00:00
abstract：:The most commonly used method for specifying the locations of functional areas in the human cerebral cortex is the coordinate system of Talairach and Tournoux (Co-planar Stereotaxic Altas of The Human Brain (1988) Georg Thieme Verlag, Stuttgart). It was designed to locate subcortical nuclei by reference to an axis joi...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(03)00046-3
更新日期：2003-05-30 00:00:00
abstract：:In this study, we developed an immuno-autoradiographic method to obtain quantitative estimates of tyrosine hydroxylase (TH) protein in tissue sections from post-mortem human brain. Protein from tissue sections containing the locus coeruleus (LC) was directly transferred to a polyvinylidene fluoride (PVDF) membrane. Im...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(00)00211-9
更新日期：2000-06-30 00:00:00
abstract：:Sources of stereotaxic variability in long-tailed macaques (Macaca fascicularis) were examined by X-ray techniques. Accuracy in the vertical dimension can be improved significantly by measuring from the top of the brain rather than from the Frankfurt plane established by external bony landmarks. Even greater accuracy ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(85)90029-9
更新日期：1985-04-01 00:00:00
abstract：:We developed a high speed voice coil based whisker stimulator that delivers precise deflections of a single whisker or group of whiskers in a repeatable manner. The device is miniature, quiet, and inexpensive to build. Multiple stimulators fit together for independent stimulation of four or more whiskers. The system c...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2010.12.008
更新日期：2011-03-15 00:00:00
abstract：:Recently, resting state functional connectivity (RSFC) studies based on fMRI and EEG/MEG have provided valuable insight into the intrinsic functional architecture of the human brain. However, whether functional near infrared spectroscopy (fNIRS), a suitable imaging method for infant and patient populations, can be use...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2009.11.010
更新日期：2010-02-15 00:00:00
abstract：:Nitric oxide (NO) is a gas messenger with diverse physiological roles in the nervous system, from modulation of synaptic plasticity and neurogenesis to the mediation of neuronal death. NO production in the brain is catalyzed by three isoforms of NO synthase (NOS) including neuronal NOS (nNOS), inducible NOS and endoth...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2009.02.006
更新日期：2009-05-15 00:00:00
abstract：:Corticomuscular coherence (CMC) estimation is a frequency domain method used to detect a linear coupling between rhythmic activity recorded from sensorimotor cortex (EEG or MEG) and the electromyogram (EMG) of active muscles. In motor neuroscience, rectification of the surface EMG is a common pre-processing step prior...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2011.11.001
更新日期：2012-03-30 00:00:00
abstract：:Skeletal muscle is a convenient model system for studying basic questions on the neural regulation of synaptogenesis and of many properties of sarcolemma and contractile apparatus. The study of the neural signals involved in a particular regulation and of the mediating intracellular pathways, requires the chronic appl...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(00)00352-6
更新日期：2001-04-30 00:00:00
abstract：:We have developed a quantitative reverse transcription polymerase chain reaction (RT-PCR) method, using both internal and external standardisation, to quantitate fg amounts of pre-pro-thyrotropin releasing hormone (pre-pro-TRH) mRNA in the paraventricular nucleus (PVN) of individual laboratory rats. A constant amount ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(96)00090-8
更新日期：1996-10-01 00:00:00
abstract：:A microfluidic chip for culturing neurons and spatially isolating axons from somas is presented for use with visually guided whole-cell electrophysiological measurements. A modular design consisting of detachable and re-sealable layers is used to satisfy the requirements of both long-term neuron culturing as well as e...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2012.10.013
更新日期：2013-01-30 00:00:00
abstract：:A method which improves carbon fiber microelectrode selectivity for cationic amines by electrocoating the fiber with a thin film of the ionic polymer, Nafion, is described. The selectivity and response speed of these electrodes for the detection of electroactive cationic and anionic species found in brain extracellula...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(87)90011-2
更新日期：1987-12-01 00:00:00
abstract：:Capsule application of Diamidino Yellow (DY) to the cut end of the sciatic nerve immediately followed by capsule application of Fast Blue (FB) resulted in approximately 95% double-labelled dorsal root ganglion neurones (DRGn) and motoneurones (Mn). Nerve injection of DY followed either immediately or 2 months later by...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(99)00164-8
更新日期：2000-02-15 00:00:00
abstract：:We assessed the potential of using multielectrode arrays (MEAs) to investigate several physiological properties of the calyx of Held synapse in the medial nucleus of the trapezoid body of gerbil. Due to the large size of the synapse, it became widely employed in studies on synaptic mechanisms. Electrical stimulation a...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2008.07.015
更新日期：2008-09-30 00:00:00
abstract：:This paper describes a new method for the analysis of ensemble coding in populations of receptor afferents. The method is based on principal component analysis (PCA) combined with algorithms for calculation of stimulus separation (discrimination). It allows a quantification of the ability of ensembles of afferents to ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(95)00089-5
更新日期：1995-12-01 00:00:00
abstract：:Depression is characterized by a lack of "motivation" rather than a lack of "physical space" to move around. This study was designed to evaluate predictivity of an open space swimming test for antidepressant activity of various antidepressants in rats. Without drug treatment, rats showed a significant reduction in the...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(03)00068-2
更新日期：2003-06-15 00:00:00
abstract：BACKGROUND:While several novel therapeutic approaches for HD are in development, resources to conduct clinical trials are limited. Large animal models have been proposed to improve assessment of safety, tolerability and especially to increase translational reliability of efficacy signals obtained in preclinical studies...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2015.11.013
更新日期：2016-05-30 00:00:00
abstract：:I describe here the successful visualization of interhemispheric callosal connections in the live mammalian cortex. The development of this method was prompted by the finding that fluorescent tracer labeling of groups of cortical neurons, when done under optimal conditions, is sufficiently intense to be visible even i...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(88)90137-9
更新日期：1988-10-01 00:00:00
abstract：:A rat model of penetrating ballistic-like brain injury (PBBI) was recently established to study military-relevant severe traumatic brain injury (TBI). The purpose of this study was to conduct a side-by-side evaluation of two well-established cognitive testing paradigms: the novel object recognition (NOR) task and the ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2010.03.012
更新日期：2010-05-30 00:00:00
abstract：:The study aim was to develop a model to generate cerebral evoked potentials (CEPs) by mechanical distention of the duodenum in rats. Twenty Sprague-Dawley rats were anaesthetized and the EEG recorded from the left and right somatosensory cortices (S1L, S1R). A balloon catheter was implanted into the duodenum. A pneuma...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2007.01.014
更新日期：2007-05-15 00:00:00
abstract：:We investigated the usefulness of the Overall Rotarod Performance (ORP) test for evaluating overall locomotory ability in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-injected-mouse model of Parkinson's disease (PD). For this procedure, the mice are pretrained on the rotarod and then tested at a series of i...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(98)00078-8
更新日期：1998-09-01 00:00:00
abstract：:Intracranial self-stimulation (ICSS) in the rat is a useful tool for studying the importance of various brain monoamines in positive reinforcement. The effects of compounds interacting with dopaminergic neurotransmission is measurable by studying the changes of reward thresholds. By computerisation of the analysis of ...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(93)90084-5
更新日期：1993-05-01 00:00:00
abstract：:Implantable microfabricated microelectrode arrays represent a versatile and powerful tool to record electrophysiological activity across multiple spatial locations in the brain. Spikes and field potentials, however, correspond to only a fraction of the physiological information available at the neural interface. In ur...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2008.06.036
更新日期：2008-09-15 00:00:00
abstract：:The present study describes the use of nose-poke habituation as a memory task and demonstrates that it is sensitive to hypo- and hypermnestic pharmacological treatments administered post-trial. Habituation of nose-poke behavior of rats was defined as a reduction in number of nose-pokes compared to baseline. It was mea...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/0165-0270(94)00143-5
更新日期：1995-03-01 00:00:00
abstract：:The thalamocortical slice is widely employed for in vitro studies of cortical circuits. This preparation was developed in order to preserve anatomical and functional connectivity between the ventrobasal thalamus and somatosensory (whisker/barrel) cortex of young mice, and thalamocortical slice experiments have contrib...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/s0165-0270(02)00150-4
更新日期：2002-09-15 00:00:00
abstract：:This research evaluated changes in surface electromyographic (SEMG) spike shape across different levels of isometric force. Ninety-six subjects generated three 5-s isometric step contractions of the elbow flexors at 40, 60, 80, and 100% of maximal voluntary contraction (MVC). Force and bipolar SEMG activity were monit...
journal_title：Journal of neuroscience methods
pub_type： 杂志文章
doi：10.1016/j.jneumeth.2006.07.004
更新日期：2007-01-15 00:00:00
